LOCUS       sequence-337020-        4539 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The plasmids contain inserts (ORFs) for expressing various members 
            of the GlycoLipid Transfer Protein (GLTP) superfamily..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4539)
  AUTHORS   Simanshu DK, Kamlekar RK, Wijesinghe DS, Zou X, Zhai X, Mishra SK, 
            Molotkovsky JG, Malinina L, Hinchcliffe EH, Chalfant CE, Brown RE, 
            Patel DJ
  TITLE     Non-vesicular trafficking by a ceramide-1-phosphate transfer protein
            regulates eicosanoids.
  JOURNAL   Nature. 2013 Aug 22;500(7463):463-7. doi: 10.1038/nature12332. Epub 
            2013 Jul 17.
  PUBMED    23863933
REFERENCE   2  (bases 1 to 4539)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nature12332"; journalName: "Nature"; date: "2013-08-22- 
            22"; volume: "500"; issue: "7463"; pages: "463-7"
COMMENT     Sequence Label: pSUMO CPTP
FEATURES             Location/Qualifiers
     source          1..4539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     544..563
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(583..660)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     706..725
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        706..724
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    725..749
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             805..822
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             832..849
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             862..1155
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     regulatory      1382..1391
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1798..1842
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1880..1898)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1894..1941
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2171..2986)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2171..2986)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2894..2913
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2987..3078)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3094..3843)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(3104..3121)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3344..3363)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4039..4539,1..582))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
       61 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
      121 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
      181 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
      241 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
      301 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
      361 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
      421 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
      481 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
      541 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
      601 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
      661 cgggatctcg acgctctccc ttatgcgact cctgcattag gaaattaata cgactcacta
      721 taggggaatt gtgagcggat aacaattccc ctgtagaaat aattttgttt aactttaata
      781 aggagatata ccatgggcag cagccatcat catcatcatc acagcagcgg cctggtgccg
      841 cgcggcagcc acatggcaag catgtcggac tcagaagtca atcaagaagc taagccagag
      901 gtcaagccag aagtcaagcc tgagactcac atcaatttaa aggtgtccga tggatcttca
      961 gagatcttct tcaagatcaa aaagaccact cctttaagaa ggctgatgga agcgttcgct
     1021 aaaagacagg gtaaggaaat ggactcctta agattcttgt acgacggtat tagaatccaa
     1081 gctgatcaga cccctgaaga tttggacatg gaggataacg atattattga ggctcacaga
     1141 gaacagattg gtggatccgg tttcaatctg aaagtcgtcc tggtcagttt caagcagtgt
     1201 ctcgatgaga aggaagaggt cttgctggac ccctacattg ccagctggaa gggcctggtc
     1261 aggtttctga acagcctggg caccatcttc tcattcatct ccaaggacgt ggtctccaag
     1321 ctgcggatca tggagcgcct caggggcggc ccgcagagcg agcactaccg cagcctgcag
     1381 gccatggtgg cccatgagct gagcaaccgg ctggtggacc tggagcgccg ctcccaccac
     1441 ccggagtctg gctgccggac ggtgctgcgc ctgcaccgcg ccctgcactg gctgcagctg
     1501 ttcctggagg gcctgcgtac cagccccgag gacgcacgca cctccgcgct ctgcgccgac
     1561 tcctacaacg cctcgctggc cgcctaccac ccctgggttg tgcgccgcgc cgtcaccgtg
     1621 gccttctgca cgctgcccac acgcgaggtc ttcctggagg ccatgaacgt ggggcccccg
     1681 gagcaggccg tgcagatgct aggcgaggcc ctccccttca tccagcgtgt ctacaacgtc
     1741 tcccagaagc tctacgccga gcactccctg ctggacctgc cctaggtcga gtctggtaaa
     1801 gaaaccgctg ctgcgaaatt tgaacgccag cacatggact cgtctactag cgcagcttaa
     1861 ttaacctagg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     1921 cgggtcttga ggggtttttt gctgaaacct caggcatttg agaagcacac ggtcacactg
     1981 cttccggtag tcaataaacc ggtaaaccag caatagacat aagcggctat ttaacgaccc
     2041 tgccctgaac cgacgacaag ctgacgaccg ggtctccgca agtggcactt ttcggggaaa
     2101 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2161 gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     2221 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     2281 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     2341 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     2401 gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt
     2461 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     2521 ttcgtgattg cgcctgagcg agacgaaata cgcggtcgct gttaaaagga caattacaaa
     2581 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     2641 aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta
     2701 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     2761 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     2821 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
     2881 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     2941 ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcatactc ttcctttttc
     3001 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3061 tttagaaaaa taaacaaata ggcatgcagc gctcttccgc ttcctcgctc actgactcgc
     3121 tacgctcggt cgttcgactg cggcgagcgg tgtcagctca ctcaaaagcg gtaatacggt
     3181 tatccacaga atcaggggat aaagccggaa agaacatgtg agcaaaaagc aaagcaccgg
     3241 aagaagccaa cgccgcaggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3301 aatcgacgct caagccagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3361 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3421 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg ttggtatctc
     3481 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3541 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3601 tcgccactgg cagcagccat tggtaactga tttagaggac tttgtcttga agttatgcac
     3661 ctgttaaggc taaactgaaa gaacagattt tggtgagtgc ggtcctccaa cccacttacc
     3721 ttggttcaaa gagttggtag ctcagcgaac cttgagaaaa ccaccgttgg tagcggtggt
     3781 ttttctttat ttatgagatg atgaatcaat cggtctatca agtcaacgaa cagctattcc
     3841 gttactctag atttcagtgc aatttatctc ttcaaatgta gcacctgaag tcagccccat
     3901 acgatataag ttgtaattct catgttagtc atgccccgcg cccaccggaa ggagctgact
     3961 gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt
     4021 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4081 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
     4141 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
     4201 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
     4261 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
     4321 gtccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
     4381 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
     4441 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
     4501 gcgagtgaga tatttatgcc agccagccag acgcagacg
//
